Engineering innervated secretory epithelial organoids by magnetic three-dimensional bioprinting for stimulating epithelial growth in salivary glands
Engineering innervated secretory epithelial organoids by magnetic three-dimensional bioprinting for stimulating epithelial growth in salivary glands
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DOI：
10.1016/j.biomaterials.2018.06.011
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发表时间：
2018-10-01
期刊：
影响因子：
14
通讯作者：
Ferreira, Joao N.
中科院分区：
文献类型：
--
作者：
Adine, Christabella;Ng, Kiaw K.;Ferreira, Joao N.
由于唾液腺（SG）中存在受损的分泌上皮和神经，目前基于唾液的刺激疗法对放疗引起的口干症并不完全有效。因此，三维生物工程类器官对于再生受损的SG至关重要。在这里，一个最近验证的三维（3D）生物制造系统，磁性3D生物打印（M3DB），被测试从神经嵴来源的间充质干细胞，人牙髓干细胞（hDPSC）产生神经支配的分泌上皮类器官。细胞用磁性纳米颗粒（MNP）标记，并在空间上用磁性点排列，以生成三维球体。接下来，用成纤维细胞生长因子10 （4-400 ng/ml）完成SG上皮分化阶段，以概括SG上皮的形态发生和神经发生。然后将SG类器官移植到离体模型中，观察其上皮生长和神经支配情况。与不含mnp的球体相比，m3db形成的球体具有较高的细胞存活率（>90%）和稳定的ATP胞内活性。分化后，球形细胞表达SG上皮区室，包括分泌上皮、导管上皮、肌上皮和神经元上皮。制备的类器官在FGF10刺激下也产生唾液α -淀粉酶，不同神经递质刺激可引起细胞内钙动员和跨上皮抵抗。移植后，SG样器官显著刺激受损SG的上皮和神经元生长。这是第一次生物打印具有生物功能的神经元样器官。因此，这是SG再生和治疗放疗性口干症的重要一步。(C) 2018 Elsevier Ltd.版权所有。
Current saliva-based stimulation therapies for radiotherapy-induced xerostomia are not fully effective due to the presence of damaged secretory epithelia and nerves in the salivary gland (SG). Hence, three-dimensional bio-engineered organoids are essential to regenerate the damaged SG.Herein, a recently validated three-dimensional (3D) biofabrication system, the magnetic 3D bio-printing (M3DB), is tested to generate innervated secretory epithelial organoids from a neural crest derived mesenchymal stem cell, the human dental pulp stem cell (hDPSC). Cells are tagged with magnetic nanoparticles (MNP) and spatially arranged with magnet dots to generate 3D spheroids. Next, a SG epithelial differentiation stage was completed with fibroblast growth factor 10 (4-400 ng/ml) to recapitulate SG epithelial morphogenesis and neurogenesis. The SG organoids were then transplanted into ex vivo model to evaluate their epithelial growth and innervation.M3DB-formed spheroids exhibited both high cell viability rate (>90%) and stable ATP intracellular activity compared to MNP-free spheroids. After differentiation, spheroids expressed SG epithelial compartments including secretory epithelial, ductal, myoepithelial, and neuronal. Fabricated organoids also produced salivary alpha-amylase upon FGF10 stimulation, and intracellular calcium mobilization and trans epithelial resistance was elicited upon neurostimulation with different neurotransmitters. After transplantation, the SG-like organoids significantly stimulated epithelial and neuronal growth in damaged SG.It is the first time bio-functional innervated SG-like organoids are bioprinted. Thus, this is an important step towards SG regeneration and the treatment of radiotherapy-induced xerostomia. (C) 2018 Elsevier Ltd. All rights reserved.